CN110004031A - Centrifugal classification bioreactor and micro-imaging method for separating - Google Patents
Centrifugal classification bioreactor and micro-imaging method for separating Download PDFInfo
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- 238000003384 imaging method Methods 0.000 title claims abstract description 28
- 239000007788 liquid Substances 0.000 claims abstract description 37
- 230000002572 peristaltic effect Effects 0.000 claims abstract description 35
- 238000005119 centrifugation Methods 0.000 claims abstract description 30
- 238000000926 separation method Methods 0.000 claims abstract description 22
- 239000004615 ingredient Substances 0.000 claims abstract description 11
- 241000009298 Trigla lyra Species 0.000 claims abstract description 3
- 239000000126 substance Substances 0.000 claims abstract description 3
- NJPPVKZQTLUDBO-UHFFFAOYSA-N novaluron Chemical compound C1=C(Cl)C(OC(F)(F)C(OC(F)(F)F)F)=CC=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F NJPPVKZQTLUDBO-UHFFFAOYSA-N 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 10
- 210000002381 plasma Anatomy 0.000 claims description 10
- 230000008719 thickening Effects 0.000 claims description 9
- 239000002699 waste material Substances 0.000 claims description 9
- 239000003381 stabilizer Substances 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 4
- 238000012790 confirmation Methods 0.000 claims description 3
- 239000012530 fluid Substances 0.000 claims description 3
- 238000010926 purge Methods 0.000 claims description 3
- 230000001360 synchronised effect Effects 0.000 claims description 3
- 210000004369 blood Anatomy 0.000 claims description 2
- 239000008280 blood Substances 0.000 claims description 2
- 238000011017 operating method Methods 0.000 claims description 2
- 239000002002 slurry Substances 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 1
- 238000012856 packing Methods 0.000 abstract 1
- 238000010586 diagram Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 108010019670 Chimeric Antigen Receptors Proteins 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/32—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of substances in solution
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
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Abstract
The present invention discloses a kind of centrifugal classification bioreactor and micro-imaging method for separating, the centrifugal classification bioreactor, including charging system, centrifugal separation system and imaging separation system；Charging system includes peristaltic pump, bubble inductor, several liquid storing bags and pipe clip valve；Centrifugal separation system includes centrifugation motor and rotatable reactor tank；It is centrifuged motor driven reactor tank and generates centrifugal force around center axis thereof high speed rotation, by substance centrifugation layering in reactor tank；The imaging separation system includes miniature video camera, stepper motor, separating pipe, peristaltic pump, bubble inductor, liquid storing bag and pipe clip valve；The imaging separation system can acquire the liquid layered situation in reactor tank in real time；It is compared and is imaged by control system, separating pipe rotation is driven by stepper motor, the switch of peristaltic pump and pipe clip valve, collects the required ingredient in layering in control system control piper.The present invention directly realizes centrifugation layering and packing in the bioreactor, easy to operate, avoids pollution.
Description
Technical field
The present invention relates to centrifugal classification bioreactor and micro-imaging method for separating, belong to biopharmaceutical technology,
It is related to the preparation of Chimeric antigen receptor T cell.
Background technique
Bioreactor is the container for cultivating cell or tissue, and current bioreactor passes through not yet transfers realization certainly
Centrifuge separation, it needs to be transferred in another centrifuge separation container, it is troublesome in poeration, and there is pollution in transfer process, it occupies
Equipment increases cleaning and disinfection cost and equipment investment.
Summary of the invention
To solve the above problems, the invention discloses centrifugal classification bioreactors.Present invention simultaneously provides
A kind of micro-imaging method for separating.
The specific technical solution of the present invention is as follows:
Centrifugal classification bioreactor, including charging system, centrifugal separation system and imaging separation system；
The charging system includes peristaltic pump, bubble inductor, several liquid storing bags and pipe clip valve；
The centrifugal separation system includes centrifugation motor and rotatable reactor tank；It is centrifuged motor driven reactor tank and surrounds its center
Axis high speed rotation generates centrifugal force, by substance centrifugation layering in reactor tank；
The imaging separation system includes miniature video camera, stepper motor, separating pipe, peristaltic pump, bubble inductor, liquid storing bag and pipe
Press from both sides valve；
The imaging separation system can acquire the liquid layered situation in reactor tank in real time；It is compared and is imaged by control system,
The switch of peristaltic pump and pipe clip valve in control piper.
It is radially reduced between tank mouth at the top of the reactor tank, forms horizontal shoulder, the microcam is located at platform
Above shoulder, the microcam acquires the hierarchical information in reactor tank straight down.
It is imaged in separation system, separating pipe deviates the setting of reactor tank center, and reactor tank and stepping electricity are stretched out in separating pipe upper end
Machine transmission connection, the L-shaped bending in separating pipe lower end, stepper motor drive separating pipe to surround center axis rotation, separating pipe nose end
Face is concentric radian with reactor bottle wall.
The reactor tank bottom level, and reactor tank bottom outside is provided with thickening pedestal, the thickening pedestal offers
The cross recess of right-angled intersection, the intersection point of cross recess are located at the center for thickening pedestal；
The lower contact for thickening pedestal is provided with rotary-tray, and the upper surface of the rotary-tray is equipped with the ten of right-angled intersection
Word raised line, the intersection point of cross raised line are located at the center of rotary-tray, and the cross raised line, which just coincide, to be stuck in cross recess；
Bracket is provided with below the reactor tank, the centrifugation motor is located in bracket, the central actuating shaft of motor and rotation
The center of pallet connects, and bearing, the motor driven rotary-tray rotation, rotation are provided between the rotary-tray and bracket
Pallet and reactor tank synchronous rotary.
The bearing includes bearing inner race and bearing outer ring, and is arranged in several between bearing inner race and bearing outer ring
A ball, the bearing outer ring are fixedly connected with bracket, are not contacted with rotary-tray, bearing inner race and the fixed company of rotary-tray
It connects, is not contacted with bracket.
The cover is connected with tank mouth by rotary joint, and when reactor tank rotates, cover is remained stationary.
Fixed cover is additionally provided with above the cover, fixed cover even circumferential is provided with several stabilizers,
The fixed upper backup pad of the fixed cover horizontal connection, the edge of the upper backup pad is provided with the side extended straight down
Plate, lower end and the bracket of the side plate are fixed；
The fixed cover and the equal even circumferential of cover offer several connecting holes, the connecting hole and cover of the fixed cover
Connecting hole sequentially coaxially, be provided through manual bolt in each connecting hole.
Micro-imaging centrifugal classification method, this method is based on above-mentioned centrifugal classification bioreactor, in the centrifugation
It is carried out in sorting bioreactor, including following operating procedure:
Step 1: charging: opening the corresponding peristaltic pump of Feed lines and pipe clip valve, and the blood plasma in liquid storing bag is added in tank, when adding
Bubble inductor in expects pipe road detects bubble, stops charging；After preferred blood plasma is added in tank, PBS solution is squeezed into
In blood plasma liquid storing bag, plasma bags are cleaned, addition step is repeated and continuously adds the PBS solution after cleaning in tank；It is then turned on point
It selects liquid storage bag to correspond to the peristaltic pump in pipeline and pipe clip valve, a certain proportion of sorting liquid is added in tank；
Step 2: component centrifugation: starting centrifugation motor, centrifugation motor drive reactor tank to rotate around its longitudinal center axis, centrifugation
After motor speed reaches setting value, each ingredient in component forms concentric loop shape in reactor tank, in the axial direction of reactor tank
Direction, the consistency of thickness of each layer；
Step: 3: confirmation layering is completed: layered image is acquired by microcam straight down on shoulder, then it is micro- at
As compared with preset layered image, confirming that the boundary of each layer is clear in centrifugal classification system, that is, it is judged as that layering is completed；
Step 4: starting Fen Xi mechanism: stepper motor starting drives separating pipe to move in a circle, when the nozzle of the L section of separating pipe
When touching innermost layer, start peristaltic pump, separating pipe starts to aspirate；Peristaltic pump and pipe clip valve in corresponding waste liquid collecting bag pipeline
It can open, the liquid of suction is pumped into waste liquid collecting bag；
Step 5: collect useful component layer: when separating pipe rotation is drawn into useful component layer fastly, system will by imaging contrast
Automatically switch the switch of corresponding peristaltic pump and pipe clip valve, collects useful component into liquid storing bag；When useful component has been collected, other
When layer is unwanted contributions layer, stop motor, reactor tank is stopped rotating, and falls on tank bottom under the ingredient whole in reactor tank at this time；
Step 6: discharge unwanted contributions: the liquid fallen into reactor tank will be pumped into waste fluid bag by outlet tube by peristaltic pump,
(reactor tank preferably, can be pumped into again in PBS solution purge tank, and be discharged by outlet tube).
Step 7: the peristaltic pump reversely rotated in useful component pipeline being controlled by control system, opens the pipe in corresponding pipeline
Valve is pressed from both sides, useful component is transmitted back in tank again.
Level where the useful component be it is pre-set by computer, according to the level set, system can be from
It is dynamic to open and the peristaltic pump and pipe clip valve in handoff response pipeline.
The working principle of the invention is:
The present invention starts the motor below bioreactor, motor driven bioreactor tank body high speed rotation, bioreactor
In suspension be layered by centrifugation, form circumference successively adjacent several layers in reactor tank, pass through microcam acquisition point
Layer information, whether clear according to layering abutted surface to determine whether centrifugation is complete, after complete centrifugation, layering is completed, and holding turns
Speed is constant, starts stepper motor rotating separation pipe, successively aspirates different layering ingredients from inside to outside, will after centrifugal classification
The useful component being collected into is transmitted back to again in clean tank.
The beneficial effects of the present invention are:
The present invention realizes centrifugation in the bioreactor, does not need replacement container, and the time is shorter, operates totally-enclosed full-automatic.
Detailed description of the invention
Fig. 1 is structural schematic diagram of the invention (also make an abstract attached drawing)；
Fig. 2 is the cross-sectional view of Fig. 1, i.e., assorting room schematic diagram of the present invention；
Appended drawing reference: 1-reactor tank, 2-layerings, 3-covers, 4-fixed covers, 5-connecting holes, 6-subpackage bags, 7-pipes
Folder valve, 8-microcams, 9-separating pipes, 10-cross recess, 11-cross raised lines, 12-brackets, 13-motors,
14-bearings, 15-thicken pedestal, 16-rotary-trays, 17-stabilizers.
Specific embodiment
With reference to the accompanying drawings and detailed description, the present invention is furture elucidated.It should be understood that following specific embodiments are only
For illustrating the present invention rather than limiting the scope of the invention.
Embodiment one:
Centrifugal classification bioreactor of the present invention mainly includes two parts, and one is centrifugal separation system, the other is imaging
Separation system；First by centrifugation, by the cultured products centrifugation layering 2 in bioreactor, ingredient is separated, then by independent point
The ingredient draws opened are to different subpackage bags 6.In order to which when heterogeneity layering 2, the layering 2 of same layer is being reacted in centrifugal process
The circumference surrounding consistency of thickness of tank 1,1 vertical display of reactor tank, tank mouth is upward, and tank bottom is downward, and tank bottom, which is provided with, can drive instead
Tank 1 is answered to surround the driving mechanism of center axis thereof rotation.Overall structure is referring to Fig. 1.
Embodiment two:
Further devise the mechanism for realizing centrifugal rotation are as follows: be located at the motor 13 of 1 lower section of reactor tank, motor 13 is located at bracket 12
Interior, the central actuating shaft of motor 13 is connect with the center of rotary-tray 16, is provided with bearing between rotary-tray 16 and bracket 12
14, bearing 14 includes 14 outer ring of 14 inner ring of bearing and bearing, and if being arranged between 14 outer ring of 14 inner ring of bearing and bearing
Dry ball, 14 outer ring of bearing are fixedly connected with bracket 12, do not contact with rotary-tray 16,14 inner ring of bearing and rotary-tray 16
It is fixedly connected, is not contacted with bracket 12；Motor 13 drives rotary-tray 16 to rotate, rotary-tray 16 and 1 synchronous rotary of reactor tank.
Embodiment three:
The bindiny mechanism of reactor tank 1 Yu rotary-tray 16 is devised, further in order to improve the company of reactor tank 1 Yu rotary-tray 16
It connects stability, 1 bottom level of reactor tank, and 1 bottom outside of reactor tank and is provided with and thicken pedestal 15, thicken pedestal 15 and offer ten
The cross recess 10 that word intersects, the intersection point of cross recess 10 are located at the center for thickening pedestal 15；Thicken the lower contact of pedestal 15
It is provided with rotary-tray 16, the upper surface of rotary-tray 16 is equipped with the cross raised line 11 of right-angled intersection, the intersection point of cross raised line 11
Positioned at the center of rotary-tray 16, cross raised line 11, which just coincide, to be stuck in cross recess 10.Pass through cross raised line 11 and cross
The mutual engomphosis relation of groove 10.Realize that reactor tank 1 and rotary-tray 16 can be stably connected in rotary course.
Example IV:
Further devise cover 3 is stably connected with mechanism, and in order to reinforce the stability of cover 3, the top of the cover 3 is also set
It is equipped with fixed cover 4, the surrounding of cover 3 is encased outside the surrounding of fixed cover 4,4 even circumferential of fixed cover is provided with several
Stabilizer 17.Stabilizer 17 is used to stablize the pressure of fixed cover 4.
Fixed cover 4 and the equal even circumferential of cover 3 offer several connecting holes 5, the connecting hole 5 and tank of fixed cover 4
The connecting hole 5 of lid 3 sequentially coaxially, is provided through Aerodynamic Axis in each connecting hole 5, the surrounding and connecting hole 5 of Aerodynamic Axis are close
Envelope.Aerodynamic Axis plays fixed function, and fixed cover 4 is fixedly connected with cover 3.
Upper backup pad can also be compacted on fixed cover 4, upper backup pad is connected to the side of bracket 12 by side plate,
Cover 3 is stabilized.
Fixed cover 4 and the equal even circumferential of cover 3 offer several connecting holes 5, the connecting hole 5 and tank of fixed cover 4
The connecting hole 5 of lid 3 sequentially coaxially, is provided through manual bolt in each connecting hole 5.
Embodiment five
The form of tank mouth is further devised, tank mouth reduces, and forms horizontal shoulder, micro-imaging centrifugal classification system includes position
Microcam 8 above shoulder, microcam 8 acquire 2 information of layering in reactor tank 1 straight down.By miniature
The acquisition information of video camera 8 judges whether centrifugation is complete, when 2 ingredient of layering is sucked out, when starts to inhale next layer, be switched to next
A subpackage bag 6.
Embodiment six:
Further devise Fen Xi mechanism, Fen Xi mechanism includes several subpackage bags 6 and point that is mounted on 1 cover 3 of reactor tank
From pipe, separating pipe 9 and subpackage bag 6 on multichannel hose line and are provided with 7 He of pipe clip valve by the sterile connection of multichannel hose
The work of peristaltic pump, pipe clip valve 7 and peristaltic pump is automatically controlled by a computer by imaging system.Referring to fig. 2, inlet faces
Every layer of liquid to be inhaled, separating pipe 9 are mounted on cover 3, separating pipe rotation are driven by stepper motor 13, towards reactor tank 1
Inner wall is mobile, and separating pipe 9 is mounted on the position that cover 3 deviates the center of circle.
Embodiment seven:
Micro-imaging centrifugal classification system is further devised, micro-imaging centrifugal classification system being capable of instantaneous acquiring reactor tank 1
In liquid layered 2(liquid layered 2 be followed successively by supernatant, leucocyte, red blood cell from inside to outside) situation；Micro-imaging centrifugation point
System and pipe clip valve and peristaltic pump cascade Mach-Zehnder interferometer are selected, the switch of each pipe clip valve and peristaltic pump can be controlled respectively, is carried out different
The switching of subpackage bag 6.
Embodiment eight:
The method of the present invention includes: step 1: charging: the corresponding peristaltic pump of Feed lines and pipe clip valve is opened, by the blood in liquid storing bag
Slurry is added in tank, when the bubble inductor in Feed lines detects bubble, stopping charging；Preferred blood plasma is added in tank
Afterwards, PBS solution is squeezed into blood plasma liquid storing bag, cleans plasma bags, repeated addition step and continue to add by the PBS solution after cleaning
Enter in tank；It is then turned on sorting liquid storage bag and correspond to peristaltic pump and pipe clip valve in pipeline, it will be in a certain proportion of sorting liquid addition tank；
Step 2: component centrifugation: starting centrifugation motor, centrifugation motor drive reactor tank to rotate around its longitudinal center axis, centrifugation
After motor speed reaches setting value, each ingredient in component forms concentric loop shape in reactor tank, in the axial direction of reactor tank
Direction, the consistency of thickness of each layer；
Step 3: confirmation layering is completed: layered image is acquired by microcam straight down on shoulder, then it is micro- at
As compared with preset layered image, confirming that the boundary of each layer is clear in centrifugal classification system, that is, it is judged as that layering is completed；
Step 3: starting Fen Xi mechanism: stepper motor starting drives separating pipe to move in a circle, when the nozzle of the L section of separating pipe
When touching innermost layer, start peristaltic pump, separating pipe starts to aspirate；Peristaltic pump and pipe clip valve in corresponding waste liquid collecting bag pipeline
It can open, the liquid of suction is pumped into waste liquid collecting bag；
Step 5: collect useful component layer: when separating pipe rotation is drawn into useful component layer fastly, system will by imaging contrast
Automatically switch the switch of corresponding peristaltic pump and pipe clip valve, collects useful component into liquid storing bag；When useful component has been collected, other
When layer is unwanted contributions layer, stop motor, reactor tank is stopped rotating, and falls on tank bottom under the ingredient whole in reactor tank at this time；
Step 6: discharge unwanted contributions: the liquid fallen into reactor tank will be pumped into waste fluid bag by outlet tube by peristaltic pump,
Reactor tank preferably, can be pumped into again in PBS solution purge tank, and be discharged by outlet tube；
Step 7: the peristaltic pump reversely rotated in useful component pipeline being controlled by control system, opens the pipe clamp in corresponding pipeline
Useful component is transmitted back in tank by valve again.
The technical means disclosed in the embodiments of the present invention is not limited to the technical means disclosed in the above technical means, and further includes
Technical solution consisting of any combination of the above technical features.
Taking the above-mentioned ideal embodiment according to the present invention as inspiration, through the above description, relevant staff is complete
Various changes and amendments can be carried out without departing from the scope of the technological thought of the present invention' entirely.The technology of this invention
Property range is not limited to the contents of the specification, it is necessary to which the technical scope thereof is determined according to the scope of the claim.
Claims (10)
1. centrifugal classification bioreactor, which is characterized in that including charging system, centrifugal separation system and imaging separation system；
The charging system includes peristaltic pump, bubble inductor, several liquid storing bags and pipe clip valve；
The centrifugal separation system includes centrifugation motor and rotatable reactor tank；It is centrifuged motor driven reactor tank and surrounds its center
Axis high speed rotation generates centrifugal force, by substance centrifugation layering in reactor tank；
The imaging separation system includes miniature video camera, stepper motor, separating pipe, peristaltic pump, bubble inductor, liquid storing bag and pipe
Press from both sides valve；
The imaging separation system can acquire the liquid layered situation in reactor tank in real time；It is compared and is imaged by control system,
The rotation of the switch and separating pipe of peristaltic pump and pipe clip valve in control piper.
2. centrifugal classification bioreactor according to claim 1, which is characterized in that at the top of the reactor tank with tank mouth it
Between radially reduce, form horizontal shoulder, the microcam is located above shoulder, and the microcam is adopted straight down
Collect the hierarchical information in reactor tank.
3. centrifugal classification bioreactor according to claim 2, which is characterized in that in imaging separation system, separating pipe
Deviate the setting of reactor tank center, reactor tank is stretched out in separating pipe upper end and stepper motor is sequentially connected, and separating pipe lower end is L-shaped curved
Song, stepper motor drive separating pipe to surround center axis rotation, and separating pipe nozzle end face is concentric radian with reactor bottle wall.
4. centrifugal classification bioreactor according to claim 1 to 3, which is characterized in that the reactor tank bottom water
It is flat, and reactor tank bottom outside is provided with thickening pedestal, the thickening pedestal offers the cross recess of right-angled intersection, and cross is recessed
The intersection point of slot is located at the center for thickening pedestal；
The lower contact for thickening pedestal is provided with rotary-tray, and the upper surface of the rotary-tray is equipped with the ten of right-angled intersection
Word raised line, the intersection point of cross raised line are located at the center of rotary-tray, and the cross raised line, which just coincide, to be stuck in cross recess.
5. centrifugal classification bioreactor according to claim 4, which is characterized in that be provided with below the reactor tank
Bracket, the centrifugation motor are located in bracket, and the central actuating shaft of motor and the center of rotary-tray connect, the rotary-tray
Bearing, the motor driven rotary-tray rotation, rotary-tray and reactor tank synchronous rotary are provided between bracket.
6. centrifugal classification bioreactor according to claim 5, which is characterized in that the bearing include bearing inner race and
Bearing outer ring, and several balls being arranged between bearing inner race and bearing outer ring, the bearing outer ring are fixed with bracket
Connection, does not contact, bearing inner race is fixedly connected with rotary-tray, is not contacted with bracket with rotary-tray；The cover and tank mouth
It is connected by rotary joint, when reactor tank rotates, cover is remained stationary.
7. centrifugal classification bioreactor according to claim 6, which is characterized in that be additionally provided with above the cover
Fixed cover, fixed cover even circumferential are provided with several stabilizers；
The fixed upper backup pad of the fixed cover horizontal connection, the edge of the upper backup pad is provided with the side extended straight down
Plate, lower end and the bracket of the side plate are fixed；
The fixed cover and the equal even circumferential of cover offer several connecting holes, the connecting hole and cover of the fixed cover
Connecting hole sequentially coaxially, be provided through manual bolt in each connecting hole.
8. a kind of micro-imaging centrifugal classification method, this method based on above-mentioned centrifugal classification bioreactor, it is described from
It is carried out in heart sorting bioreactor, including following operating procedure:
Step 1: charging: opening the corresponding peristaltic pump of Feed lines and pipe clip valve, and the blood plasma in liquid storing bag is added in tank, when adding
Bubble inductor in expects pipe road detects bubble, stops charging；
Step 2: component centrifugation: starting centrifugation motor, centrifugation motor drive reactor tank to rotate around its longitudinal center axis, centrifugation
After motor speed reaches setting value, each ingredient in component forms concentric loop shape in reactor tank, in the axial direction of reactor tank
Direction, the consistency of thickness of each layer；
Step: 3: confirmation layering is completed: layered image is acquired by microcam straight down on shoulder, then it is micro- at
As compared with preset layered image, confirming that the boundary of each layer is clear in centrifugal classification system, that is, it is judged as that layering is completed；
Step 4: starting Fen Xi mechanism: stepper motor starting drives separating pipe to move in a circle, when the nozzle of the L section of separating pipe
When touching innermost layer, start peristaltic pump, separating pipe starts to aspirate；Peristaltic pump and pipe clip valve in corresponding waste liquid collecting bag pipeline
It can open, the liquid of suction is pumped into waste liquid collecting bag；
Step 5: collect useful component layer: when separating pipe rotation is drawn into useful component layer fastly, system will by imaging contrast
Automatically switch the switch of corresponding peristaltic pump and pipe clip valve, collects useful component into liquid storing bag；When useful component has been collected, other
When layer is unwanted contributions layer, stop motor, reactor tank is stopped rotating, and falls on tank bottom under the ingredient whole in reactor tank at this time；
Step 6: discharge unwanted contributions: the liquid fallen into reactor tank will be pumped into waste fluid bag by outlet tube by peristaltic pump,
Reactor tank preferably, can be pumped into again in PBS solution purge tank, and be discharged by outlet tube；
Step 7: the peristaltic pump reversely rotated in useful component pipeline being controlled by control system, opens the pipe clamp in corresponding pipeline
Useful component is transmitted back in tank by valve again.
9. micro-imaging centrifugal classification method according to claim 8, which is characterized in that the useful component layer is by counting
Calculation machine is pre-set, and according to the level set, system can be automatically turned on and the peristaltic pump and pipe in handoff response pipeline
Press from both sides valve.
10. micro-imaging centrifugal classification method according to claim 8 or claim 9, which is characterized in that in step 1, preferred blood
After slurry is added in tank, PBS solution is squeezed into blood plasma liquid storing bag, cleans plasma bags, repeating addition step will be after cleaning
PBS solution continuously adds in tank.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910342085.4A CN110004031B (en) | 2019-04-26 | Centrifugal sorting bioreactor and microscopic imaging sorting method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910342085.4A CN110004031B (en) | 2019-04-26 | Centrifugal sorting bioreactor and microscopic imaging sorting method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110004031A true CN110004031A (en) | 2019-07-12 |
| CN110004031B CN110004031B (en) | 2024-06-21 |
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|---|---|---|---|---|
| US4710161A (en) * | 1985-04-22 | 1987-12-01 | The Green Cross Corporation | Continuous type centrifugal separator |
| US20080041772A1 (en) * | 2006-08-17 | 2008-02-21 | Gambro Bct, Inc. | Blood Processing Apparatus with Robust Outflow Process Control |
| CN107267389A (en) * | 2017-08-23 | 2017-10-20 | 湖南开启时代生物科技有限责任公司 | A kind of cell expansion apparatus |
| CN210085426U (en) * | 2019-04-26 | 2020-02-18 | 南京比瑞生物科技有限公司 | Centrifugal sorting bioreactor |
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| US4710161A (en) * | 1985-04-22 | 1987-12-01 | The Green Cross Corporation | Continuous type centrifugal separator |
| US20080041772A1 (en) * | 2006-08-17 | 2008-02-21 | Gambro Bct, Inc. | Blood Processing Apparatus with Robust Outflow Process Control |
| CN107267389A (en) * | 2017-08-23 | 2017-10-20 | 湖南开启时代生物科技有限责任公司 | A kind of cell expansion apparatus |
| CN210085426U (en) * | 2019-04-26 | 2020-02-18 | 南京比瑞生物科技有限公司 | Centrifugal sorting bioreactor |
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